Determination of total polyphenolic compounds and flavonoids in Matricaria chamomella flowers.

Matricaria chamomella is of medicinal importance and has been used for its various pharmacological activities against different diseases due to the presence of polyphenolic compounds especially flavonoids and their glycosides. Flowers of the chamomile plant were studied for the quantification of total polyphenolic compounds and flavonoids aglycon. Total polyphenolic compounds obtained were 83.22mg as gallic acid equivalents/g of plant material; while for the determination of flavonoids aglycon the plant material was subjected to acid hydrolysis before quantification through HPLC-PDA. Results showed that luteolin, quercetin, apigenin, isorhamnetin and kaempferol were quantified. Apigenin was found in highest concentration (0.071mg/ml) while amounts of the rest of the flavonoids quantified are: luteolin (0.012mg/ml), quercetin (0.032mg/ml), kaempferol (0.001mg/ml) and isorhamnetin (0.023 mg/ml).

Quantification of polyphenolic compounds and flavonoids in Achillea millefolium and Equisetum arvense.

Flavonoids have been of considerable importance and interest because of their medicinal activity. Responding to their numerous health benefits, a comparative study on the quantitative determination of total polyphenolic compounds and flavonoids was carried out in Achillea millefolium and Equisetum arvense. Total polyphenolic compounds were quantified by Folin-Ciocalteau method using different solvents in order to prove their extraction efficiency. Focus within total polyphenolic quantification study was placed on the traditional reflux and solvents used were: water, 100% acetone, 100% ethanol, 80% ethanol, 50% methanol and 70% methanol. In order to make flavonoids free from glycosidic moiety for quantification, hydrolysis was performed in 50% MeOH at 90°C using 6 M HCl concentration. Reverse phase high performance liquid chromatography (RP-HPLC) in gradient elution mode at 50°C using Hypersil BDS (RP-18) column was employed for the separation of flavonoids. Mobile phase used consisted of different combinations of water-methanol-tetrahydrofuran-phosphoric acid. Flavonoids quantified were luteolin, quercetin, apigenin, isorhamnetin and kaempferol.

Determination of total polyphenolic compounds and flavonoids in Juglans regia leaves.

Juglans regia leaves have been widely used in traditional medicines because of its antimicrobial, antihelmintic, astringent, keratolytic, antidiarrhoeal, hypoglycaemic, depurative, tonic, carminative activity. Total polyphenolic compounds were determined using the Folin-Ciocalteau method and flavonoids were quantified using the HPLC-PDA after the hydrolysis of the plant material with HCl. Among the flavonoids myricetin, quercetin, apigenin and kaempferol were found in appreciable amount.

Quality control of herbs: determination of amino acids in Althaea officinalis, Matricaria chamomilla and Taraxacum officinale.

Analysis of raw materials and final products need reliable methods for the standardization of natural product drugs. Legal guideline also emphasizes on the qualitative and quantitative analyses of the plant constituents in an herbal product. In this study, thin layer chromatography (TLC) and amino acid analyzer was used for the determination of amino acids in plant extracts. Samples for this study were standards and aqueous extracts from Althaea officinalis, Matricaria chamomilla and Taraxacum officinale. Different amino acids in the extracts were detected through TLC. An automatic amino acid analyzer was used for the quantification of amino acids in the plant extracts under study.

Proteomics profiling of microdissected low- and high-grade prostate tumors identifies Lamin A as a discriminatory biomarker.

Proteomics screening methods for the identification of diagnostic and prognostic biomarkers in cancer are still lagging behind DNA- or RNA-based analysis. We used two-dimensional differential gel electrophoresis (2D-DIGE) in combination with laser capture microdissection (LCM) and MALDI-TOF/TOF mass spectrometry to determine differentially abundant proteins and candidate biomarkers in prostate cancer. Paired (benign and tumor) samples were isolated from 23 Gleason Score 6 (GS 6) and 23 Gleason Score 8 and higher (GS 8+) radical prostatectomy specimens and subjected to 2D-DIGE analysis. Minimal fluorescent dye labeling was applied and electrophoresis performed with triple samples (paired benign and tumor; internal control) for each case of tumor. Nineteen differently abundant proteins were identified by mass spectrometry and further validated. One half of them were associated with glycolysis and the Warburg effect; these were upregulated in tumors. The upregulation correlated with tumor dedifferentiation and might be relevant for selection of therapeutic strategies. Among the other proteins, heat shock protein 60 (HSP60) was significantly upregulated in tumor tissue compared to its benign counterpart. Furthermore, lamin A was statistically highly discriminatory between low and high Gleason score tumors and might serve as a new biomarker of tumor differentiation and prognosis.

Proanthocyanidins: target compounds as antibacterial agents.

Grape seeds accumulate in huge quantities as byproduct during wine production and are therefore a cheap source for pharmacologically active agents. However, studies prove poor antibacterial activity, and results of analyses are sometimes contradictory. The aim of this study was, thus, to determine the antibacterial activity of grape seed extracts with special focus on the chromatographic characterization of active fractions. In the course of these investigations, extraction protocols were optimized so that microwave-assisted extraction (MAE) guaranteed highest preconcentration efficiency. Proanthocyanidins, monomeric flavonoid aglycones, as well as some of their glycosides could be identified within yielded extracts via high-performance liquid chromatography-mass spectrometry (HPLC-MS). By that means the coherence number of possible isomers of procyanidins was approximated by a newly developed equation. As far as antibacterial activity determined via screening tests is concerned, the extracts generally have been found to be positively responsive toward 10 different gram-positive and gram-negative bacteria strains. After fractionation of the raw extracts, proanthocyanidins P2, P3, P4 and gallate esters P2G and P3G (P = proanthocyanidin consisting of catechin and epicatechin units, n = oligomerization degree, G = gallate ester) were determined as active antibacterial agents toward 10 different pathogens. Only moderate activity was found for monomeric flavonoid fractions.

Quality assessment and quantitative analysis of flavonoids from tea samples of different origins by HPLC-DAD-ESI-MS.

Components of green tea ( Camellia sinensis) have been of considerable interest in recent years because of their potential utility as pharmaceutical agents, particularly for their antioxidant and anticarcinogenic activity. Responding to the increasing scientific validation of numerous health benefits of tea, a comprehensive approach was adopted to carry out analysis for the quality assessment of flavonoids in tea samples of different origins. For this purpose, extraction, separation, and mass spectrometric parameters were optimized. Extraction methods evaluated include reflux extraction, a modified accelerated solvent extraction (ASE), namely, Aquasolv extraction, and microwave-assisted extraction (MAE) using different percentages of solvents. Separation was performed by a specifically developed reversed phase high-performance liquid chromatography (RP-HPLC) method using different C18 and C8 stationary phases. Optimization of extraction techniques clearly proved the performance of MAE, which delivered highest yields in a very short time. Additionally, the comparison with Aquasolv extraction provided new insights, as variations in quantified amounts of target compounds between the extracts could be explained on the basis of thermal degradation and epimerization phenomena. Especially the epimerization phenomenon for catechin/epicatechin oligomers, that is, of procyanidins P 2 and P 3, was observed for the first time. Finally, an optimized extraction and separation system was used for qualitative and quantitative investigations of compounds from different green tea samples from Ceylon (cultivated under biologically controlled conditions), Japan, India, and China as well as from one black tea sample from India.